Compositions and methods for treating mammals with modified alginates and modified pectins

ABSTRACT

A modified alginate and/or modified pectin composition and method for preventing and/or treating diseases and/or conditions caused by circulating agents such as poisonous heavy metals, environmental toxins, calcium and cholesterol, is provided. The composition includes a modified alginate having a molecular weight of no more than 40,000 daltons and/or a modified pectin having a molecular weight of no more than 40,000 daltons. The method involves orally or intravenously administering the modified alginate and/or modified pectin composition, alone or with excipient.

This application is a Continuation of application Ser. No. 10/199,009,filed Jul. 22, 2002, which is a Continuation of application Ser. No.09/883,329, filed Jun. 19, 2001, now U.S. Pat. No. 6,462,029, which is aContinuation-In-Part of application Ser. No. 09/255,265, filed Feb. 23,1999, now U.S. Pat. No. 6,274,566, the entireties of which are hereinincorporated by reference.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention generally relates to low molecular weight alginateand/or pectin compositions and methods of using the same. The lowmolecular weight alginate and/or pectin compositions, which bind tovarious agents, including disease agents, heavy metals and othersubstances, protect mammals from and treat mammals with diseases andconditions. The low molecular weight alginate and/or pectin compositionsare particularly useful in removal of deleterious agents or deleteriousamounts of indigenous agents from a patient's blood supply and aretherefore useful in heavy metal and environmental toxin removal as wellas in calcium and cholesterol removal for treatment of arteriosclerosis.

2. Background of the Technology

It is well known that gelling agents may be effective in binding to avariety of substances, in vitro. While high molecular weight bindingsubstances are known, due to their size, it is difficult for suchsubstances to pass various blood barriers, including the gut or tocirculate freely in the bloodstream. This is also, in part, becausetheir solubility in water is generally too low.

Cancer metastasis is a stage of cancer which frequently marks conversionto an incurable disease. Most cancers share the ability to metastasize.In the metastatic process, cancer cells depart from the primary tumor,invade the basement membrane, traverse the bloodstream as tumor cellemboli and interact with the vascular endothelium of the target organ.There, the cancer cells proliferate to form secondary colonies. Kohn,Anti-Cancer Res., 13:2553-2259 (1993). It has been demonstrated thatoral administration of modified citrus pectin, i.e., low molecularweight pectin, may be effective in suppressing or preventing metastases,by binding to residues on cancer cell surfaces, presumably throughinteraction between galactosyl residues on the modified pectin withsurface galectin-3, a carbohydrate binding protein on the surface ofcancer cells. Pienta, et al., J. Nat. Cancer Inst. 87, 348-353 (1995).It has also been demonstrated in application Ser. No. 09/255,265 thatlike modified pectin, modified alginate is effective in preventingand/or treating the metastatic process.

Cancer cells are not the only inimical elements circulating in mammalianblood. In many cases, diseases and/or conditions are mediated bycirculating inorganic and/or organic agents. Thus, deleterious agents oreven increased amounts of indigenous substances can induce such diseasesor conditions. For example, calcium has been associated with a number ofdisease conditions, including arteriosclerosis and calcinosis.Arteriosclerosis is also influenced by the presence of cholesterol inthe bloodstream.

While there are a variety of treatments for removal of detrimentalamounts of circulating agents such as heavy metals, typically, theseembrace removal, and treatment of portions of a patient's blood supplyand reintroduction to the patient.

Algin and pectin, or alignates or pectinates digestion products of each,are known gelling agents. Algins and pectins occur naturally; a chiefsource of algins is seaweed; a chief source of pectins is citrus.Alginates are formed by a mixture of polymannuronic acid andpolyguluronic acid. Repeating sub-units are bound by glycosidic links atthe 1a-4a-di-axial position (polyguluronic acid) while repeatingsub-units of polymannuronic acid are bound by galactic links at the1e-4e di-equatorial position. Pectins are polymers of galacturonic acidthat may be partially esterified. Naturally occurring algins andnaturally occurring pectins are high molecular weight products. Bothcan, however, be reduced to low molecular weight products (definedherein as 40,000 daltons or less) by either chemical treatment (e.g.,alkaline hydrolysis) or enzymatic degradation.

Alginates generally have superior protein recovery rates when comparedto pectins of relatively similar molecular weight. Clare, IndustrialGums (1993 3^(rd) Edition). Alginates therefore may exhibit a strongerbinding ability than pectins for many substances.

Besides binding cancer cells, alginates are known to be effective metalbinding compounds. This, along with the superior recovery rateassociated with alginates, lends support that alginates would be usefulin the prevention and/or treatment of diseases and/or conditions.

U.S. Pat. No. 5,141,927 discloses the use of an alginate and vitamin Dcomposition to treat hypertension.

U.S. Pat. No. 5,597,810 discloses the use of an alginate-cholesterolcompound to attract undesired lipids in the fluid media of the digestivetract prior to their absorption across cells lining the tract.

U.S. Pat. Nos. 5,283,086 and 5,324,526 disclose an algin-containing foodor beverage as a source of dietary fiber. The patents disclose that thealgin-containing food or beverage, which contains an aqueous solution ofan algin having a weight average molecular weight in the range fromabout 10,000 to about 150,000, can be used to suppress the absorption ofharmful substances in the digestive tract.

Japanese Patent No. 09235234 (Abstract) discloses the use of alginateoligosaccharides in the intestine to treat hyperlipidemia andarteriosclerosis.

Kimura et al., J. Ethnopharmacol, 54(1):47-54 (1996) (Abstract),discloses a study of natural sodium alginate (AG-270 (i.e., 2,700 kDa))and three low-molecular weight sodium alginates (AG-1 (10 kDa), AG-5 (50kDa) and AG-10 (100 kDa)) on cholesterol excretion and glucose tolerancein rats. Kimura et al. discloses that the study revealed that thenatural sodium alginate and two of the three modified sodium alginatesenhanced cholesterol secretion and inhibited blood glucose levels fromrising after glucose administration. The authors noted that the resultssuggest that the effects of natural sodium alginate and AG-5 and AG-10on cholesterol excretion and glucose tolerance may be due to inhibitionof cholesterol and glucose adsorption from the small intestine bygelling of free alginic acid converted in the stomach and that thesealginates can be used in the treatment/prevention of obesity,hypercholesterolemia and diabetes.

None of the references discussed above disclose substances that binddeleterious agents or deleterious amounts of indigenous agents in thebloodstream. In particular, none of the references discussed abovedisclose substances that are able to be absorbed through the intestinalmucosa, enter the bloodstream and exert their effects. In addition, noneof the references discussed above disclose chelation and removal ofheavy metals. Accordingly, there is a need to provide a composition andmethod by which disease and/or conditions caused by deleterious agentsor deleterious amounts of indigenous agents circulating in thebloodstream can be prevented or reduced.

It is an object of the invention to provide a safe and effectivetreatment for high circulating amounts of detrimental agents such asheavy metals and environmental toxins.

It is an additional object of the invention to provide a safe andeffective treatment for high circulating amounts of indigenous agentssuch as calcium and other minerals which contribute deposits such ascalcinosis (large calcium deposits present in auto-immune condition likedermatomyositis) as well as cholesterol in order to prevent and or treatatherosclerosis. A further object is to provide a method for treatingarteriosclerosis with reduced side effects.

SUMMARY OF THE INVENTION

The above objects, and other objects that are made clear by thediscussion below, are met by the oral or the intravenous administrationof modified alginates, modified pectins, or a combination of both, aloneor together with promoter compounds.

For the prevention and/or treatment of diseases and/or conditions,modified alginates and/or modified pectins are employed herein. Modifiedalginates and modified pectins refer to low molecular weight products,i.e., no more than 40,000 daltons, obtained by hydrolysis or enzymaticdigestion of algin or pectin.

Low molecular weight alginates and/or low molecular weight pectins canbe used in chelation therapy, that is, administration to bind heavymetals, toxins and calcium in the bloodstream. While these potentialpoisons (including mercury, lead, arsenic, radioactive materials andothers) can be chelated in the intestine, the inventive composition andmethod are particularly effective to chelate heavy metals circulating inthe bloodstream. The chelation of calcium, as well as cholesterolbinding, is effective in the treatment of arteriosclerosis.

Methods for modifying pectins to obtain low molecular weight pectins areknown to those of skill in the art and can be obtained from commercialsources. See, e.g., Pienta, et al. Modified alginates are produced by asimilar method, either through alkaline hydrolysis or enzymaticdegradation using alginate lyase. The final modified alginate or pectinmust be water soluble and have a weight average molecular weight of40,000 daltons or below. Preferably, the modified alginate or modifiedpectin has a weight average molecular weight between about 10,000daltons and about 20,000 daltons. More preferably, the modified alginateor modified pectin has a weight average molecular weight of about 10,000daltons. Also, preferably, the modified alginate or modified pectin hasa degree of esterification of less than about 10%. It may be combinedwith pharmaceutically acceptable carriers suitable for oral orintravenous administration, depending on the treatment method desired.Dosage levels will vary from 5-1500 mg per kg of body weight, per day,and may be sustained over a prolonged period. A preferred range may be10 mg/kg/day to 1,000 mg/kg/day.

The invention itself together with further objects and advantages, willbest be understood by reference to the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

For the treatment or prevention of diseases and/or conditions, modifiedalginates and/or modified pectins are used, either alone, or incombination. In other methods of treatment described herein, modifiedalginates or modified pectins, or their combination, may effectively beused.

A water soluble, low molecular weight modified alginate is prepared fromalginic acid. An aqueous suspension is prepared, and the pH is adjustedto 10.0 with the addition of a hydroxide, suitably sodium hydroxide, for30 minutes. This is followed by a reduction to a pH of 3.0 by theaddition of HCl, according to the method of Albersheim. et al., Arch.Biochem. Biophys., 90:46-49 (1960). pH modification is conducted attemperatures of 85-95° C. for a period of up to 24-48 hours. Afterthermal degradation over an interval lasting 8-48 hours, sodiumhydroxide and/or potassium hydroxide are added to equilibrate themodified alginate to a pH of approximately 6.0-6.5. The process may bemonitored throughout the thermal degradation stage, to evaluatemolecular weight. The essential characteristics of the inventivemodified alginate is that it be water soluble and of a molecular weightbelow 40,000 daltons. This is necessary to ensure adequate passage ofthe modified alginate to the blood during digestion. The modifiedalginate may be dried by drum or spray drying.

In an alternate, more preferred process, the alginic acid is degradedenzymatically, rather than thermally. In this process, an aqueouspreparation of alginic acid is modified to a pH of 3.0-6.0, with anoptimum pH value of about 4.5, using a mixture of sodium and potassiumhydroxide. The concentration of alginic acid in the aqueous preparationfor both thermal and enzymatic degradation is approximately 10-30%. Inthe enzymatic degradation, the temperature for maximum enzymaticactivity is 30-40° C., and is maintained over a period of 5-7 days. Asthe case with thermal degradation, the enzymatic degradation process canbe monitored to evaluate the molecular weight. The enzymatic process,which permits attainment of more accurate molecular weight andesterification of final product, allows for better standardization,better quality control and reproducible results. (Molecular weightherein is weight average molecular weight). pH is adjusted to a value of6.0-6.5, and the solution is dried by drum or spray drying.

A principle method of using the modified alginate of the invention is inthe prevention or treatment of diseases and/or conditions caused bydeleterious circulating agents. Because the individual units of modifiedalginates, guluronic acid and mannuronic acid, can be linked andcross-linked in a variety of different fashions (and obtained that wayfrom a variety of different seaweeds) the resulting units can bepresented in a variety of concentrations and accessibilities.Preferably, a large variety of modified alginate is employed to maximizethe likelihood that effective binding between the modified alginate anddeleterious circulating agents in the bloodstream can be achieved. It isbelieved that certain modified alginate configurations, includingreduced cross-linking, and a high degree of alternating monomers mayresult in the exposure of greater binding units, providing for greatereffectiveness.

In accordance with an embodiment of the invention, modified alginateand/or modified pectin is administered to chelate poisonous metals,including mercury, lead, nickel and others, that may be circulating inthe blood. Blood poisoning due to the presence of heavy metals andrelated toxins can be treated by administration of modified alginateand/or modified pectin to bind the discreet metal particles. In thisrespect, modified alginate and modified pectin are gelling agents, andthe bound particles/gel are safely removed by the body. Dosage levelsfor this type of treatment can be on the order of 50-1,000 mg ofmodified alginate and/or modified pectin per kg body weight. It shouldbe noted that many poisonous metals enter the bloodstream throughaccidental or unintentional ingestion, pollution and ingestion ofamalgam from silver fillings and other metals during dental work andfrom leakage. The modified alginate or modified pectin of the inventionis of sufficient molecular weight and is water-soluble to be readilyabsorbed through the intestinal mucosa into the bloodstream. Oraladministration of modified alginate and/or modified pectin in thismethod effectively isolates and removes dangerous metals whether presentintestinally or in the bloodstream. Metals such as aluminum, antimony,beryllium, bismuth, mercury, platinum, thallium, thorium, tin, tungsten,uranium, lead, mercury, cadmium, arsenic, silver, nickel, radioactivemetals and mixtures thereof, which (1) are frequently concentrated byplants and animals that constitute part of the food chain or (2) areotherwise present in the environment and (3) are recognized toxins thatmay effectively be removed by this method. In addition, additionaltoxins such as MTBA and dioxin may also be removed.

The present invention will be further illustrated by the followingnon-limiting Example.

EXAMPLE

The modified alginate and/or modified pectin may be employed to bindheavy metals in the blood stream as follows:

Patients that are known to have increased levels of toxic metals (e.g.,lead, mercury, cadmium, etc.) as determined through physical exams,specific questionnaires and hair analysis are chosen. Patients havingheavy metal toxicity are entered into the study.

Modified alginate and/or modified pectin with or without excipient isadministered to bind heavy metals in the blood stream and facilitateheavy metal excretion via the urinary tract. For example, modifiedalginate and/or modified pectin powder encapsulated into gelatincapsules in the amount of 800 mg/capsule may be employed. Alternatively,a water-based preparation could be used. Dosage may be 6 capsules takenthree times a day with 8 full ounces of water or juice.

An open labeled, non-controlled study with prescreened patients known tohave heavy metal burdens that exceed the normal range is conducted. Tenpre-screened patients (ages 18-75) are monitored under semi-controlledconditions using questionnaires, and are evaluated for heavy metaltoxicity at baseline, and again at one week and three week time pointsin the study. All samples are taken under the control of a qualifiedmedical professional.

Heavy metal burden is measured at time zero and at one and three weeksrespectively to determine if the oral administration of modifiedalginate and/or modified pectin (<40,000 daltons) (1) is absorbed intothe blood stream, (2) is bound with heavy metals and (3) is able tofacilitate heavy metal excretion into the urine.

The modified alginate and/or modified pectin may be administered orallyor intravenously. Alginates and pectins have been shown to be welltolerated through both methods of administration. A daily dosage ofabout 15 grams of modified alginate is believed to be effective, butdosages can vary from 5 mg of modified alginate per kg of body weight onup to 1,000-1,500 mg of modified alginate per kg of body weight, andmore, per day.

Protocol

1) Pre-qualified patients will be measured for heavy metal toxicityburden using standard urine analysis by a qualified FDA approvedlaboratory.

2) Analysis will establish the baseline levels and mid and end-pointlevels for a variety of heavy metals including but not limited to Pb⁺²,Hg⁺², Cd⁺² and As.

3) Patients will be enrolled in an opened labeled, non-controlled trialand supplemented with a novel form of modified alginate and/or modifiedpectin (<40,000 daltons). Dosages will be based on 800 mg capsules to betaken as six (6) capsules three times a day (total=14.4 grams) of themodified alginate and/or modified pectin product.

4) All patients will receive a three week's supply of alginate and/ormodified pectin capsules at a time and compliance will be measured by aquestionnaire and the honor system.

5) A qualified professional will collect urine samples at time 0(baseline) and again at one week and three-week time points. The studywill be concluded after three weeks and all data will be collected andanalyzed.

Evaluations

1) All data will be collected at the start of the study and at one weekand the endpoint at 3 weeks (21 days).

2) A FDA certified medical laboratory would process all urine samplesusing ICP-MS technology.

3) Results on toxic metal levels in the urine will be reported as mcg/gcreatine and as mcg/24 hour urine. These will account for urine dilutionvariations that may occur and are a standard measure of toxic burden andmonitoring of toxic detoxification.

4) Analytical results will be compiled and compared against baselinevalues using appropriate statistical methods.

Patients that show a statistically significant change in various toxic 5elements will be indicative of the absorption and binding of heavymetals via the bloodstream. Increased levels of heavy metals asreflected in urine samples will demonstrate the ability of modifiedalginates to specifically:

1) Be absorbed into the blood stream following oral ingestion.

2) Bind with various heavy metals circulating in the bloodstream, and

3) Facilitate the removal of heavy metals via the urinary tract.

The protocol discussed above can also be used to determine the effectsof the modified alginate and/or modified pectin (and any excipient) onadditional deleterious circulating agents, e.g., environmental toxins,and deleterious amounts of indigenous agents.

The modified alginate and/or modified pectin may be administered alone,or together with agents which enhance binding, such as whey protein(rich in glutathione), selenium and related binding adjuvants.Additional effective agents are those which aid excretion through boththe kidneys and intestines, such as diuretics, phase two initiators suchas sulfured amino acids, EDTA, etc.

Neither modified alginate nor modified pectin is known to have any sideeffects, or to exhibit cytopathology or toxicology. Oral administrationcan be a reconstitution of dried modified alginate and/or modifiedpectin in water or other suitable solution, or using a wide variety ofconventional excipients, vehicles, flavorings and the like. Additionalingredients such as phase II detoxification enhancers, botanicals,anti-oxidants, sulfured amino acids, EDTA, curcumin, indoles such as13C, selenium, zinc and glutathione, may be used to potentiate theeffects of the modified alginate and/or modified pectin composition.Higher molecular weight align and/or pectin may also be added to themodified alginate or modified pectin composition to enhance binding ofsubstances in the intestine. Controlled dosage formulations arepreferred to ensure adequate medication over time.

In this respect, the administration of modified alginate may be doublyeffective in the treatment of diseases characterized by an inadequateimmune response. As noted previously, the modified alginate of theinvention is comprised of guluronic and mannuronic acid monomers.Unbound mannuronic acid is broken down by the body to uronic acid andmannose. It is known that the administration of mannose stimulates theimmune system. Turner, Scand. J. Immunol., 48 (2):124-126 (1998);Kakkanaiah. et al., Clin. Diagn. Lab Immunol., 5(3):319-321 (1998). Toachieve significant immune stimulation, modified alginate levels shouldbe on the order of 10-15 grams/day.

The present inventors have also observed that the inventive modifiedalginate and/or modified pectin is capable of binding circulatingcalcium and circulating cholesterol. Effective binding of circulatingcalcium (although not all calcium) and circulating cholesterol areeffective methods for preventing and/or treating arteriosclerosis. Theinvention permits binding of calcium and cholesterol beyond theintestines. By binding these circulating indigenous agents, deposit ofplaque on the endothelial walls is prevented, thus providing aneffective method of treating arteriosclerosis and related conditions,including hypertension and coronary heart disease, without significantside effects. In the prevention and/or treatment of target individualswho may be at risk of developing severe arteriosclerosis, dosage valuesof 2-15 grams of modified alginate and/or modified pectin per day, or inthe range of 150 mg of modified alginate and/or modified pectin per kgbody weight, is appropriate.

The invention has been described by both specific example, and genericdescription. Alternatives, particularly alternatives with respect to thecomposition and characteristics of modified alginate and/or modifiedpectin, as well as treatment regimen, will occur to those of ordinaryskill in the art, without the exercise of inventive faculty. Suchalternatives remain within the scope of the invention, unlessspecifically excluded by the recitation of the claims set forth below.

1-19. (canceled)
 20. A method of treating a condition or diseasemediated by an agent selected from the group consisting of a poisonousmetal, an environmental toxin and calcium circulating in the blood, thedisease or condition being selected from the group consisting of bloodpoisoning, arteriosclerosis, atherosclerosis, calcinosis,dermatomytosis, obesity, hypercholesterolima, and diabetes, the methodcomprising administering to a mammal in need of same an effective amountof pectin comprised of partially esterified polymers of galacturonicacid having a molecular weight of no more than 40,000 daltons, in anamount to effectively bind said agent.
 21. The method of claim 20,wherein said condition is metal poisoning, and said agent is a poisonousheavy metal.
 22. The method of claim 21, wherein said heavy metal isselected from the group consisting of aluminum, antimony, beryllium,bismuth, mercury, platinum, thallium, thorium, tin, tungsten, uranium,lead, cadmium, arsenic, silver, nickel, radioactive metals and mixturesthereof.
 23. The method of claim 20, wherein said pectin is administeredorally or intravenously.